Geerlings A, Hallard D, Martinez Caballero A, Lopes Cardoso I, van der Heijden R, Verpoorte R
Leiden/Amsterdam Center for Drug Research, Division of Pharmacognosy, Leiden University, Gorlaeus Laboratories, P.O. Box 9502, 2300 RA Leiden, The Netherlands e-mail:
Plant Cell Rep. 1999 Dec;19(2):191-196. doi: 10.1007/s002990050732.
Cinchona officinalis 'Ledgeriana', former called Cinchona ledgeriana, hairy roots were initiated containing constitutive-expression constructs of cDNAs encoding the enzymes tryptophan decarboxylase (TDC) and strictosidine synthase (STR) from Catharanthus roseus, two key enzymes in terpenoid indole and quinoline alkaloid biosynthesis. The successful integration of these genes and the reporter gene gus-int was demonstrated using Southern blotting and the polymerase chain reaction. The products of TDC and STR, tryptamine and strictosidine, were found in high amounts, 1200 and 1950 μg g dry weight, respectively. Quinine and quinidine levels were found to rise up to 500 and 1000 μg g dry weight, respectively. The results show that genetic engineering with multiple genes is well possible in hairy roots of C. officinalis. However, 1 year after analyzing the hairy roots for the first time, they had completely lost their capacity to accumulate alkaloids.
金鸡纳树‘莱杰氏种’（原称金鸡纳树莱杰氏变种），其发根中导入了来自长春花的编码色氨酸脱羧酶（TDC）和异胡豆苷合酶（STR）的cDNA组成型表达构建体，这两种酶是萜类吲哚和喹啉生物碱生物合成中的关键酶。通过Southern杂交和聚合酶链反应证明了这些基因与报告基因gus-int的成功整合。发现TDC和STR的产物色胺和异胡豆苷含量很高，分别为1200和1950μg/g干重。发现奎宁和奎尼丁水平分别上升至500和1000μg/g干重。结果表明，在金鸡纳树发根中进行多基因遗传工程是完全可行的。然而，在首次分析发根一年后，它们完全丧失了积累生物碱的能力。
